摘要
目的:观察低极性人参皂苷F4(F4)抑制人单核系白血病细胞SHI-1和红系白血病细胞K562增殖及诱导分化的作用。方法:以不同浓度(0、40、80和120 mg/L)的F4处理SHI-1和K562细胞,MTT比色法和半固体集落形成实验检测F4对细胞增殖的抑制作用;瑞氏-吉姆萨染色法观察细胞趋向分化的形态;流式细胞术和免疫荧光法检测分化相关抗原CD11b和CD14阳性表达率及其荧光反应强度;免疫细胞化学和Western blot法分析SHI-1细胞分化相关转录因子PU.1和K562细胞红系分化相关GATA-1和γ-globin的蛋白表达水平;RT-qPCR法检测PU.1和GATA-1的mRNA表达变化。结果:与对照(0 mg/L)组比较,F4能够抑制SHI-1和K562细胞增殖(P<0.01),诱导白血病细胞趋向分化,表现为细胞核/质比缩小,染色质粗糙,核仁减少,可见空泡,SHI-1细胞核有扭曲。SHI-1细胞CD11b和K562细胞CD14阳性表达率显著升高(P<0.01),且免疫荧光反应从弱阳性转变为强阳性;但SHI-1细胞CD14和K562细胞CD11b阳性表达率无显著变化。同时,F4上调PU.1和GATA-1的mRNA和蛋白表达水平及γ-globin的蛋白表达水平(P<0.05)。结论:低极性人参皂苷F4具有抑制SHI-1和K562白血病细胞增殖及诱导分化的抗白血病作用,其作用可通过调控分化相关PU.1、GATA-1和γ-globin的表达来实现。
AIM:To investigate the effects of low-polarity ginsenoside F4(F4)on the proliferation and differ⁃entiation of human monocytic leukemia SHI-1 cells and erythroid leukemia K562 cells.METHODS:The SHI-1 cells and K562 cells were treated with F4 at different concentrations(0,40,80 and 120 mg/L).The suppressive effect of F4 on cell proliferation was detected by MTT assay and colony formation assay with semi-solid culture.The morphologic features of the cells were observed under inverted microscope with Wright-Giemsa staining.The positive expression rates and fluores⁃cence intensity of differentiation-associated antigens CD11b and CD14 were detected by flow cytometry and immunofluores⁃cence method.The protein expression of differentiation-related transcription factor PU.1 in SHI-1 cells,and GATA-1 andγ-globin related to erythroid differentiation in K562 cells was analyzed by immunocytochemistry and Western blot.The mRNA expression of PU.1 and GATA-1 was detected by RT-qPCR.RESULTS:Compared with control(0 mg/L)group,F4 inhibited the proliferation of SHI-1 cells and K562 cells(P<0.01),and induced these leukemic cells to differentiate,showing reduced nucleus/cytoplasm ratio,rough chromatin,reduced nucleoli,increased vacuoles,and distortion of SHI-1 cell nucleus.The positive expression rates of CD11b in SHI-1 cells and CD14 in K562 cells were increased significantly(P<0.01),and the immunofluorescence reaction changed from weak positive to strong positive.No significant change in the positive expression rates of CD14 in SHI-1 cells and CD11b in K562 cells was observed.At the same time,F4 up-regulated the mRNA and protein expression levels of PU.1 and GATA-1,and the protein level ofγ-globin(P<0.05).CON⁃CLUSION:The low-polarity ginsenoside F4 possesses anti-leukemia effect that inhibits the proliferation of SHI-1 cells and K562 cells,and induces differentiation by regulating the expression of differentiation-related PU.1,GATA-1 andγ-globin.
作者
李弯
高瑞兰
余潇苓
尹利明
唐比强
兰高琛
兰金剑
赵燕娜
LI Wan;GAO Rui-lan;YU Xiao-ling;YIN Li-ming;TANG Bi-qiang;LAN Gao-chen;LAN Jin-jian;ZHAO Yan-na(The First Affiliated Hospital of Zhejiang Chinese Medical University,Hangzhou 310006,China.)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2020年第11期1988-1995,共8页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.81774068)
浙江省自然科学基金资助项目(No.LY20H290004,No.LQ19H290002)。